Autofluorescence in mosses.

Images made through a microscope. All subject types.

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WalterD
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Autofluorescence in mosses.

Post by WalterD »

After the sporangia I wanted to jump back to mosses to observe autofluorescence. (Determination for follow up!)
The first 2 pictures are showing a moss with leafs that cannot be seen with the bare eye; looks like green velvet. Picture 1 shows a side view, 2 a top view which is more difficult to stack. The sample was found in an industrial environment.

Picture 3 shows the spore capsules of a moss with very small hair-like leafs. Noted mosses can have spore capsules althrough the year in Europe.

When taking picture 4 the photobleaching was (as always) an issue. I observed the bleaching started in the tips of the leafs and propagated to the inside. Meantime the leafs started to curl up. Within the first 20 seconds of exposure the object lost about 75% of its radiation. The same method was used as in the previous posts last 2 pictures: obscuring the mercury vapour lamp any time the camera sensor was not exposed.

Picture 5 is a better illustration of the "glow in the dark" effect of autofluorescence on a moss leaf, generated by the chlorophyl under UV excitation. The tip of that leaf is asking for a higher magnification...
1.
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2.
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3.
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4.
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5.
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All done with UV excitation, YOC-3 filter and 9x epi-objective (3,4,5), 7x (1,2).

Jacek
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Post by Jacek »

Very nice

Beatsy
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Post by Beatsy »

Interesting set. Is there such a thing as UV flash so as to reduce the photo-bleaching issue?

Sumguy01
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Post by Sumguy01 »

:smt038 Very nice.
Thanks for sharing.

Marek Mis
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Post by Marek Mis »

Very nice images Walter, especially the first one !

Marek

WalterD
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Post by WalterD »

My pleasure

Image

(Beatsy) Interesting set. Is there such a thing as UV flash so as to reduce the photo-bleaching issue?
I wish there was. The mercury arc vapour lamp needs to burn at least 2 hours each session, otherwise it will not last the 200 burning hours. There are diaphragm shutters with controllers, (https://www.thorlabs.com/newgrouppage9. ... up_id=6619) such could be integrated in the instrument between the lamp and the optics. However, besides introducing another source of vibration, it should be talking with Canon EOS Utility, which is impossible as far as I know.
Another option would be utilizing "smart glass", this is transparant material that darkens when a current is sent through it. Here the same issue: how to trigger anything that's not incorporated in the Canon software? If anybody has ideas please share it here...

Regarding the picture: sorry no ID yet, 7x/0.17 epi objective.
Zerene Dmap substacks.

rjlittlefield
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Post by rjlittlefield »

WalterD wrote:Canon EOS Utility
...
Here the same issue: how to trigger anything that's not incorporated in the Canon software? If anybody has ideas please share it here...
Consider using a keyboard/mouse "robot" to drive both Canon EOS Utility and the controller(s) for other software. Something like http://www.robot-soft.com/ .

I have worked with several people who use this approach with camera control software to acquire focus stacks.

--Rik

carlos.uruguay
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Post by carlos.uruguay »

Super!!

WalterD
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Post by WalterD »

Thanks Carlos!
Consider using a keyboard/mouse "robot" to drive both Canon EOS Utility and the controller(s) for other software. Something like http://www.robot-soft.com/ .

I have worked with several people who use this approach with camera control software to acquire focus stacks.
That's brilliant Rik! What you mentioned is an application mimicking clicking a mouse on a pre-defined location on the screen. This will enable me to use the mouse for the shutter controller simultaneously. With this knowledge it will be a matter of time before it's incorporated within my setup. Anyhow, there's room enough to install it :D
Hopefully I can beat the price barrier, what I spotted sells for about $1000...

This whole issue would be worth a separate post in the equipment discussions section, maybe I'll do that this weekend. Meantime I've created a 6-seconds animation showing the effect of photobleaching and UV light on moss as shown in picture 4 (now under postprocessing/ being retouched) https://www.youtube.com/watch?v=qrFOkjV ... e=youtu.be
Be warned the compression resulted in poor image quality, the clip is also too dark.

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